Otimização do método para a análise da enzima redutase do nitrato na cultura do milho
Autor(a) principal: | |
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Data de Publicação: | 2022 |
Tipo de documento: | Trabalho de conclusão de curso |
Idioma: | por |
Título da fonte: | Biblioteca Digital de Monografias da UFMT |
Texto Completo: | http://bdm.ufmt.br/handle/1/3981 |
Resumo: | In Brazil, corn occupies great importance and prominence among the products of the agricultural sector due to its versatile use and has prominence among grasses for the response with greater intensity to nitrogen fertilization. Nitrogen is absorbed mainly in the form of nitrate (NO3-) and this anion is first reduced to nitrite (NO2-) by the enzyme Nitrate Reductase (NR). Determining nitrate reductase activity (aRN) becomes important because plants with high enzyme activity have a higher ability to assimilate nitrogen and respond to nitrogen fertilization. Mulder, Boxma and van Veen (1959) were the first to publish the in vivo nr assay in leaf discs of olericultural plants, however, each species has different anatomical and physiological characteristics, so it is necessary to establish appropriate and particular analytical conditions for each type of plant. The general objective of this study was to standardize and characterize the optimal conditions for determining the activity of nitrate reductase enzyme (aRN) in fresh corn leaves. The BM 3051 hybrid was cultivated at Fazenda Escola Boa Esperança located in Barra do Garças - MT. To estimate aRN, the simplified in vivo method of Mulder, Boxma and van Veen (1959) was used. Diagnostic leaves of corn were collected and incubated in potassium nitrate solution at 0.25 M for two hours at 35°C. Subsequently, 1 mL of this solution was transferred to volumetric balloon and sulphanyl acid, alphanaphthylamine and sodium acetate buffer were added. After solution rested, small amount was transferred to a cuvette and read in spectrophotometer set to 540nm. The optimization of the method consisted of sequential experiments, in which different conditions of preparation the incubation medium were studied, varying: type of cut of the fresh leaf sample, leaf tissue mass used, incubation time of the samples, potassium nitrate concentration (KNO3) in the buffer solution and incubation temperature. In each experiment, one factor was modified and, for the next one, the highest aRN value observed in the previous experiment was based. The proposed changes to the current enzymatic method to be optimized in corn crop consist of cutting the samples of diagnostic leaves in the whole disk format, using 0.2 g of leaf tissue mass that should be incubated in solution with 0.16 M KNO3 in phosphate buffer pH 7.4, for 1 hour and 30 minutes, temperature that can range from 30°C to 35°C. The conditions for quantifying nitrate reductase enzyme activity in corn leaves were standardized. |
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Otimização do método para a análise da enzima redutase do nitrato na cultura do milhoCNPQ::CIENCIAS AGRARIAS::AGRONOMIAZea maysmétodo in vivopadronizaçãoatividade enzimáticaZea maysin vivo methodstandardizationenzymatic activityIn Brazil, corn occupies great importance and prominence among the products of the agricultural sector due to its versatile use and has prominence among grasses for the response with greater intensity to nitrogen fertilization. Nitrogen is absorbed mainly in the form of nitrate (NO3-) and this anion is first reduced to nitrite (NO2-) by the enzyme Nitrate Reductase (NR). Determining nitrate reductase activity (aRN) becomes important because plants with high enzyme activity have a higher ability to assimilate nitrogen and respond to nitrogen fertilization. Mulder, Boxma and van Veen (1959) were the first to publish the in vivo nr assay in leaf discs of olericultural plants, however, each species has different anatomical and physiological characteristics, so it is necessary to establish appropriate and particular analytical conditions for each type of plant. The general objective of this study was to standardize and characterize the optimal conditions for determining the activity of nitrate reductase enzyme (aRN) in fresh corn leaves. The BM 3051 hybrid was cultivated at Fazenda Escola Boa Esperança located in Barra do Garças - MT. To estimate aRN, the simplified in vivo method of Mulder, Boxma and van Veen (1959) was used. Diagnostic leaves of corn were collected and incubated in potassium nitrate solution at 0.25 M for two hours at 35°C. Subsequently, 1 mL of this solution was transferred to volumetric balloon and sulphanyl acid, alphanaphthylamine and sodium acetate buffer were added. After solution rested, small amount was transferred to a cuvette and read in spectrophotometer set to 540nm. The optimization of the method consisted of sequential experiments, in which different conditions of preparation the incubation medium were studied, varying: type of cut of the fresh leaf sample, leaf tissue mass used, incubation time of the samples, potassium nitrate concentration (KNO3) in the buffer solution and incubation temperature. In each experiment, one factor was modified and, for the next one, the highest aRN value observed in the previous experiment was based. The proposed changes to the current enzymatic method to be optimized in corn crop consist of cutting the samples of diagnostic leaves in the whole disk format, using 0.2 g of leaf tissue mass that should be incubated in solution with 0.16 M KNO3 in phosphate buffer pH 7.4, for 1 hour and 30 minutes, temperature that can range from 30°C to 35°C. The conditions for quantifying nitrate reductase enzyme activity in corn leaves were standardized.No Brasil, o milho ocupa grande importância e destaque dentre os produtos do setor agrícola decorrente de seu uso versátil e possui destaque entre as gramíneas pela resposta com maior intensidade a adubação nitrogenada. O nitrogênio é absorvido, principalmente, na forma de nitrato (NO3 - ) e esse ânion é primeiramente reduzido a nitrito (NO2 - ) pela enzima Nitrato Redutase (NR). Determinar a atividade da redutase do nitrato (aRN) se torna importante pois plantas com alta atividade da enzima tem maior capacidade de assimilar nitrogênio e de responder a adubação nitrogenada. Mulder, Boxma e van Veen (1959) foram os primeiros a publicar o ensaio in vivo da NR em discos foliares de plantas olerícolas, porém, cada espécie apresenta características anatômicas e fisiológicas diferentes, assim, é necessário se estabelecer condições analíticas apropriadas e particulares para cada tipo de planta. O objetivo geral do trabalho foi padronizar e caracterizar as condições ótimas para determinação da atividade da enzima redutase do nitrato (aRN) em folhas frescas de milho. O híbrido BM 3051 foi cultivado na Fazenda Escola Boa Esperança localizada em Barra do Garças MT. Para se estimar a aRN, utilizou-se o método in vivo simplificado de Mulder, Boxma e van Veen (1959). Foram coletadas folhas diagnósticas de milho e incubadas em solução de nitrato de potássio a 0,25 M, por duas horas, a 35°C. Posteriormente, foi transferido 1 mL desta solução parabalão volumétrico e adicionado ácido sulfanílico, alfanaftilamina e tampão de acetato de sódio. Após repouso da solução, pequena quantidade foi passada para cubeta e lida em espectrofotômetro ajustado a 540nm. A otimização do método consistiu em experimentos sequenciais, em que foram estudadas diferentes condições de preparo do meio de incubação, variando: tipo de corte da amostra de folhas frescas, massa de tecido foliar utilizada, tempo de incubação das amostras, concentração de nitrato de potássio (KNO3) na solução tampão e temperatura de incubação. Em cada experimento modificou-se um fator e, para o seguinte, tomou-se como base o maior valor de aRN observado no experimento anterior. As alterações propostas ao método enzimático atual para que seja otimizado na cultura do milho consistem em cortar as amostras de folhas diagnósticas no formato de disco inteiro, sendo utilizadas 0,2 g de massa de tecido foliar que deverão ser incubados em solução com 0,16 M de KNO3 em tampão fosfato pH 7,4, por 1 hora e 30 minutos, a temperatura que pode variar de 30°C a 35°. As condições para quantificar a atividade da enzima redutase do nitrato em folhas de milho foram padronizadas.Universidade Federal de Mato GrossoBrasilInstituto de Ciências Exatas e da Terra (ICET) – AraguaiaUFMT CUA - AraguaiaAgronomia - CUASantos, Carlos Leandro Rodrigues dos0054668076313809http://lattes.cnpq.br/0054668076313809Moraes, Milton Ferreira de1541223007968886http://lattes.cnpq.br/1541223007968886Santos, Carlos Leandro Rodrigues dos0054668076313809http://lattes.cnpq.br/0054668076313809Melo, Suzana Pereira de0392957201020352http://lattes.cnpq.br/0392957201020352Freitas, Natasha Gomes1790587653092024http://lattes.cnpq.br/1790587653092024Abreu, Gabrielly Catarine Backes2024-07-04T17:10:05Z2024-04-022024-07-04T17:10:05Z2022-12-01info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/bachelorThesisinfo:eu-repo/semantics/datasetABREU, Gabrielly Catarine Backes. Otimização do método para a análise da enzima redutase do nitrato na cultura do milho. 2022. 29 f. Monografia (Graduação em Agronomia) - Instituto de Ciências Exatas e da Terra, Universidade Federal de Mato Grosso, Barra do Garças, 2022.http://bdm.ufmt.br/handle/1/3981porinfo:eu-repo/semantics/openAccessreponame:Biblioteca Digital de Monografias da UFMTinstname:Universidade Federal de Mato Grosso (UFMT)instacron:UFMT2024-07-13T07:12:40Zoai:localhost:1/3981Biblioteca Digital de Monografiahttps://bdm.ufmt.br/PUBhttp://200.129.241.122/oai/requestopendoar:2024-07-13T07:12:40falseBiblioteca Digital de Monografiahttps://bdm.ufmt.br/PUBhttp://200.129.241.122/oai/requestbibliotecacentral@ufmt.br||opendoar:2024-07-13T07:12:40Biblioteca Digital de Monografias da UFMT - Universidade Federal de Mato Grosso (UFMT)false |
dc.title.none.fl_str_mv |
Otimização do método para a análise da enzima redutase do nitrato na cultura do milho |
title |
Otimização do método para a análise da enzima redutase do nitrato na cultura do milho |
spellingShingle |
Otimização do método para a análise da enzima redutase do nitrato na cultura do milho Abreu, Gabrielly Catarine Backes CNPQ::CIENCIAS AGRARIAS::AGRONOMIA Zea mays método in vivo padronização atividade enzimática Zea mays in vivo method standardization enzymatic activity |
title_short |
Otimização do método para a análise da enzima redutase do nitrato na cultura do milho |
title_full |
Otimização do método para a análise da enzima redutase do nitrato na cultura do milho |
title_fullStr |
Otimização do método para a análise da enzima redutase do nitrato na cultura do milho |
title_full_unstemmed |
Otimização do método para a análise da enzima redutase do nitrato na cultura do milho |
title_sort |
Otimização do método para a análise da enzima redutase do nitrato na cultura do milho |
author |
Abreu, Gabrielly Catarine Backes |
author_facet |
Abreu, Gabrielly Catarine Backes |
author_role |
author |
dc.contributor.none.fl_str_mv |
Santos, Carlos Leandro Rodrigues dos 0054668076313809 http://lattes.cnpq.br/0054668076313809 Moraes, Milton Ferreira de 1541223007968886 http://lattes.cnpq.br/1541223007968886 Santos, Carlos Leandro Rodrigues dos 0054668076313809 http://lattes.cnpq.br/0054668076313809 Melo, Suzana Pereira de 0392957201020352 http://lattes.cnpq.br/0392957201020352 Freitas, Natasha Gomes 1790587653092024 http://lattes.cnpq.br/1790587653092024 |
dc.contributor.author.fl_str_mv |
Abreu, Gabrielly Catarine Backes |
dc.subject.por.fl_str_mv |
CNPQ::CIENCIAS AGRARIAS::AGRONOMIA Zea mays método in vivo padronização atividade enzimática Zea mays in vivo method standardization enzymatic activity |
topic |
CNPQ::CIENCIAS AGRARIAS::AGRONOMIA Zea mays método in vivo padronização atividade enzimática Zea mays in vivo method standardization enzymatic activity |
description |
In Brazil, corn occupies great importance and prominence among the products of the agricultural sector due to its versatile use and has prominence among grasses for the response with greater intensity to nitrogen fertilization. Nitrogen is absorbed mainly in the form of nitrate (NO3-) and this anion is first reduced to nitrite (NO2-) by the enzyme Nitrate Reductase (NR). Determining nitrate reductase activity (aRN) becomes important because plants with high enzyme activity have a higher ability to assimilate nitrogen and respond to nitrogen fertilization. Mulder, Boxma and van Veen (1959) were the first to publish the in vivo nr assay in leaf discs of olericultural plants, however, each species has different anatomical and physiological characteristics, so it is necessary to establish appropriate and particular analytical conditions for each type of plant. The general objective of this study was to standardize and characterize the optimal conditions for determining the activity of nitrate reductase enzyme (aRN) in fresh corn leaves. The BM 3051 hybrid was cultivated at Fazenda Escola Boa Esperança located in Barra do Garças - MT. To estimate aRN, the simplified in vivo method of Mulder, Boxma and van Veen (1959) was used. Diagnostic leaves of corn were collected and incubated in potassium nitrate solution at 0.25 M for two hours at 35°C. Subsequently, 1 mL of this solution was transferred to volumetric balloon and sulphanyl acid, alphanaphthylamine and sodium acetate buffer were added. After solution rested, small amount was transferred to a cuvette and read in spectrophotometer set to 540nm. The optimization of the method consisted of sequential experiments, in which different conditions of preparation the incubation medium were studied, varying: type of cut of the fresh leaf sample, leaf tissue mass used, incubation time of the samples, potassium nitrate concentration (KNO3) in the buffer solution and incubation temperature. In each experiment, one factor was modified and, for the next one, the highest aRN value observed in the previous experiment was based. The proposed changes to the current enzymatic method to be optimized in corn crop consist of cutting the samples of diagnostic leaves in the whole disk format, using 0.2 g of leaf tissue mass that should be incubated in solution with 0.16 M KNO3 in phosphate buffer pH 7.4, for 1 hour and 30 minutes, temperature that can range from 30°C to 35°C. The conditions for quantifying nitrate reductase enzyme activity in corn leaves were standardized. |
publishDate |
2022 |
dc.date.none.fl_str_mv |
2022-12-01 2024-07-04T17:10:05Z 2024-04-02 2024-07-04T17:10:05Z |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/publishedVersion info:eu-repo/semantics/bachelorThesis info:eu-repo/semantics/dataset |
format |
bachelorThesis |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
ABREU, Gabrielly Catarine Backes. Otimização do método para a análise da enzima redutase do nitrato na cultura do milho. 2022. 29 f. Monografia (Graduação em Agronomia) - Instituto de Ciências Exatas e da Terra, Universidade Federal de Mato Grosso, Barra do Garças, 2022. http://bdm.ufmt.br/handle/1/3981 |
identifier_str_mv |
ABREU, Gabrielly Catarine Backes. Otimização do método para a análise da enzima redutase do nitrato na cultura do milho. 2022. 29 f. Monografia (Graduação em Agronomia) - Instituto de Ciências Exatas e da Terra, Universidade Federal de Mato Grosso, Barra do Garças, 2022. |
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http://bdm.ufmt.br/handle/1/3981 |
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por |
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por |
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openAccess |
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Universidade Federal de Mato Grosso Brasil Instituto de Ciências Exatas e da Terra (ICET) – Araguaia UFMT CUA - Araguaia Agronomia - CUA |
publisher.none.fl_str_mv |
Universidade Federal de Mato Grosso Brasil Instituto de Ciências Exatas e da Terra (ICET) – Araguaia UFMT CUA - Araguaia Agronomia - CUA |
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reponame:Biblioteca Digital de Monografias da UFMT instname:Universidade Federal de Mato Grosso (UFMT) instacron:UFMT |
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Biblioteca Digital de Monografias da UFMT |
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Biblioteca Digital de Monografias da UFMT |
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Biblioteca Digital de Monografias da UFMT - Universidade Federal de Mato Grosso (UFMT) |
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